A new class of cell-derived dimeric mitogens with apparently restricted specificity for vascular endothelial cells has recently been identified and generally designated vascular endothelial growth factors (VEGFs). The mitogen has been purified from: conditioned growth media of rat glioma cells, [Conn et al., Proc. Nat. Acad. Sci. USA 87: 1323-1327 (1990)]; conditioned growth media of bovine pituitary folliculo stellate cells [Ferrara and Henzel, Biochem. Biophys. Res. Comm. 161: 851-858 (1989) and Gospodarowicz et al., Proc. Natl. Acad. Sci. USA 86: 7311-7315 (1989)]. An endothelial cell growth factor isolated form mouse neuroblastoma cell line NB41 with an unreduced molecular mass of 43-51 kDa and a reduced mass of 23-29 kDa has been described by Levy et al., Growth Factors 2: 9-19 (1989). Connolly et al. (J. Biol. Chem. 264: 20017-20024 [1989]; J. Clin. Invest. 84: 1470-1478 [1989]) describe a human vascular permeability factor that stimulates vascular endothelial cells to divide in vitro and promotes the growth of new blood vessels when administered into healing rabbit bone grafts or rat corneas. An endothelial cell growth factor has been purified from the conditioned medium of the AtT-20 pituitary cell line by Plouet et al., EMBO Journal 8: 3801-3806 (1989). The growth factor was characterized as a heterodimer composed of subunits with molecular mass of 23 kDa. Leung et al. (Science 246: 1306-1309 [1989]), Keck et al. (Science 246: 1309-1312 [1989]) and Conn et al. (Proc. Natl. Acad. Sci USA 87: 2628-2632 [1990]) have described cDNAs which encode VEGF A which is homologous to the A and B chains of platelet-derived growth factor. Vascular endothelial growth factor I (VEGF I, VEGF AA) is a homodimer with an apparent molecular mass of 46 kDa, with each subunit having an apparent molecular mass of 23 kDa. VEGF I has distinct structural similarities to platelet-derived growth factor (PDGF), a mitogen for connective tissue cells but not vascular endothelial cells from large vessels.
It is, accordingly, an object of the present invention to provide novel vascular endothelial growth factor C subunit DNA free of other mammalian DNA. Another object is to provide recombinant genes capable of expressing VEGF C subunit or dimer. Another object is to provide vectors containing the DNA sequences for VEGF A or B plus C subunits. A further object is to provide a host cell transformed with a vector containing the DNA sequence for VEGF A or B plus C or VEGF C alone. It is also an object to provide a recombinant process for making VEGF C subunit. Another object is to provide a novel vascular endothelial cell growth factor which contains the C subunit. This may include heterodimers AC and BC and homodimer CC.
Vascular endothelial cell growth factor C subunit DNA is prepared by polymerase chain reaction techniques. The DNA encodes a protein that may exist as either a heterodimer or homodimer. The protein is a mammalian vascular endothelial cell mitogen and as such is useful for the promotion of vascular development and repair. This unique growth factor is also useful in the promotion of tissue repair.